Fig. 2

Effect of knockdown of I. scapularis transcripts on LGTV infection of ISE6 cells. a ISE6 cell viability following transfection with 10ng dsRNA for 60 h and 12 h post-infection (hpi) with LGTV, compared to the pGEM negative control. Results represent 2–5 technical replicates and 3 biological replicates. b Effect of I. scapularis transcript knockdown on LGTV genome replication in ISE6 cells following transfection with 10ng dsRNA for 60 h and 12 hpi with LGTV as compared to the negative control. Comparison of fold change in transcripts for the LGTV negative strand were normalized to the I. scapularis β-tubulin gene and expressed relative to percentage of pGEM control. Results represent 2 technical replicates (each with 3 machine replicates) and 2 biological replicates. c Effect of I. scapularis transcript knockdown on release of infectious LGTV from ISE6 cells transfected with dsRNA for 60 h prior to LGTV infections as assessed by plaque assay and normalized to the pGEM dsRNA negative control. Results represent 5 technical replicates and 2 biological replicates. d Effect of I. scapularis transcript knockdown on release of infectious LGTV from ISE6 cells infected with LGTV and transfected with dsRNA for 60 h. Results represent 2–5 technical replicates and 3 biological replicates. Error bars represent SEM for unpaired t-tests comparing cell viability, negative strand levels or pfu/ml of the negative pGEM control versus each gene of interest. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001. Abbreviations: FAH, fumarylacetoacetase; ERP29, endoplasmic reticulum protein 29; ALDH, aldehyde dehydrogenase; VNN, carbon-nitrogen hydrolase/vanin-like; MDH2, malate dehydrogenase; PARP, poly [ADP-ribose] polymerase; CMPK, cytidine/urdine monophosphate kinase; ACAT1, acetyl-CoA acetyltransferase; Hypo195 and Hypo576, hypothetical proteins 195 and 576; pGEM, pGEM plasmid negative control (light gray bars); LGTV 3UTR, 3’ UTR of LGTV positive control (dark gray bars); RLU, relative light units